INTERNATIONAL JOURNAL OF IMMUNOPATHOLOGY AND PHARMACOLOGY Vol. 23, no.

1,25-34 (2010)

DETAILED CHARACTERISATION OF CB2 RECEPTOR PROTEIN EXPRESSION IN

PERIPHERAL BLOOD IMMUNE CELLS FROM HEALTHY HUMAN VOLUNT1i:ERS
USING FLOW CYTOMETRY

E.S. GRAHAM!, C.E. ANGELl, L.E. SCHWARCZ!, P.R. DUNBAR2,3 and M. GLASS l

'Department ofPharmacology and Clinical Pharmacology, Faculty ofMedical and Health

Sciences, 'School ofBiological Sciences, Faculty ofScience, "Maurice Wilkins Centre, University of
Auckland, Auckland, New Zealand

Received April 6, 2009 - AcceptedAugust 7, 2009

The first two authors contributed equally to this work

It is commonly accepted from gene expression studies that the CB2 receptor is expressed by most
cell types of the rodent and human immune system. However, the exact identity of cells expressing
CB2 receptor protein in human blood or the abundance of receptors expressed by each immune subset
is not well characterised. We conducted a detailed analysis of CB2 protein levels expressed by blood-
derived immune cells from healthy human donors. Flow-cytometry was conducted using 4 commercially
available anti-CB2 polyclonal antibodies in conjunction with a selection ofimmune cell specific markers.
Across multiple healthy subjects we observed that NK cells, B-Iymphocytes and monocytes expressed a
higher level of CB2 receptor than CD4+or CD8+T-Iymphocytes. Neutrophils also expressed a low level of
CB2 receptor. NK cells had the greatest variation in CB2 expression levels, whereas for each of the other
cell types CB2 levels were relatively similar between subjects. In contrast to other methods, the high
sensitivity of flow-cytometry revealed that CB2 receptors are present on resting T-Iymphocytes at low
abundance in some healthy subjects. These data provide the first detailed analysis of CB2 protein levels
in blood leukocyte subsets from healthy donors and identifies the cell types which could be targeted with
CB2-mimetic drugs in humans.

It has been known since their discovery that both
CB1 and CB2 cannabinoid receptors are expressed
by immune cells (1-2). In recent years there has
been a resurgence of interest to identify precisely
which cells of the human immune system express
cannabinoid receptors in an effort to understand
their therapeutic potential (3-6). This is especially
relevant to CB2 as it has a more restricted localisation
(7), making it a more attractive drug target (6, 8-9).
Recent rodent studies have demonstrated that CB2
ligands can prevent inflammation-mediated tissue
injury in several inflammatory conditions (10-12).
In humans, marijuana smoking severely suppresses
alveolar macrophage activity and the production
of inflammatory cytokines (13), implicating
cannabinoid receptors in these immunosuppressive
events. However, the precise pattern of CB2 protein
expression in the human immune system is not
fully understood. Therefore, in this study we have
characterised the expression of CB2 receptor protein

Key words: cannabinoids, immune cells, leukocytes, CB2, CBl, human

Mailing address: Dr E Scott Graham,
Department of Pharmacology and Clinical Pharmacology
Faculty of Medical and Health Sciences
University of Auckland
Private Bag 92019, 1142 New Zealand
Tel: ++64 9 3737599 Fax: ++64 93737556
e-mail: s.graham@auckland.ac.nz
0394-6320 (2010)
Copyright © by BIOLIFE, s.a.s,
This publication andlor article is for individual use only and may not be further
reproduced without written permission from the copyright holder.
25 Unauthorized reproduction may result in financial and other penalties
26 E.S. GRAHAM ET AL.
by each immune cell-type in human blood from
healthy individuals to establish "basal" receptor
levels in steady-state cells. This is an important
requisite for future therapeutic CB2 strategies for
humans.
The ability to analyse CB2 receptor expression at
the protein level, has been hampered greatly by the
lack of reliable commercially available antibodies.
Therefore, most evidence for expression has relied
upon the detection of CB2 mRNA (1, 7) with some
studies using pharmacological evidence to support
the presence of functional cannabinoid receptors (9,
14-18). The mRNA for CB2 has been detected under
various conditions in most immune cell types (7, 19-
20). In the human immune system, B-lymphocytes
and natural killer cells express the highest levels
of CB2 mRNA, followed by monocytes. However,
the relative level of CB2 protein in immune cells of
healthy individuals has not been studied. Recently,
several CB2 antibodies have been validated for
detection of endogenous CB2 receptors in human
tissues (7, 15, 21-25). The objective of this study
is to conduct a detailed comparative analysis of
CB2 protein expression by each leukocyte subset in
human blood from healthy individuals using flow-
cytometry. This will enable us to identify which
human immune cells are capable of responding to
locally released endocannabinoid ligands as well as
cannabinoid-mimetic drugs. With a comprehensive
understanding of CB2 expression, we can begin
to determine the functions within each leukocyte-
subset and identify potential cannabinoid targets for
the regulation of inflammatory responses in humans
(11-12, 25- 26).
MATERIALS AND METHODS
Collection of human blood and preparation of cell
cultures
Blood samples (15-20 mL) were collected from 8
healthy volunteers after informed consent under ethical
approval from the University of Auckland human ethics
committee (reference #2009-101). Blood was collected
into EDTA-coated collection tubes and maintained at
room temperature. For the neutrophil and whole-blood
inclusion experiments direct immunofluorescence
staining was conducted following the BD Lyse/No-
Wash protocol (BD-Biosciences) using freshly collected
blood. Peripheral blood mononuclear cells (PBMC)
were prepared from whole-blood following gradient
centrifugation through Histopaque 1077 (Sigma) using
Leucosep tubes (27). Following centrifugation at 800 x
g the PBMC were recovered from the plasma layer and
washed gently several times with RPMl media. Single
cell suspensions were cryopreserved in 50% RPMI 1640,
40% FBS and 10% dimethyl sulfoxide using a protocol
which does not affect cell surface phenotype (28).
CE2 antibody panel
The CB2 antibodies used in this study were purchased
from Affinity Bioreagents (ABR, Cat # PAll-744), Sigma
(Cat # CI483), Abeam (Cat # ab3561), and Santa Cruz
(Cat # sc-l 0071). These antibodies recognise epitopes
located within the N-terminus of the CB2 protein. The
ABR CB2 antibody was raised against the first 33 amino
acids of human CB2, whereas the Abeam and Sigma CB2
antibodies were raised to residues 1-32 of rat CB2 and
cross-react with human CB2. The immunogen for the
Santa Cruz CB2 antibody is described as "mapping near
to the N-terminus of the human CB2 receptor". The exact
epitope sequence has not been declared by Santa Cruz.
Flow cytometry analysis
All flow-cytometry experiments were conducted using
a FACSCalibur flow-cytometer (Becton-Dickenson)
using standardised techniques (28-29). The following
fluorophore-conjugated mouse monoclonal antibodies
detecting CD3 [UCRTl], CD8 [SKI], CD16 [3G8] and
CD69 [L78] (BD Bioseiences, San Diego, CA); CD3
[UCRTl], CD4 [RPA-T4], CD14 [UCHM1], CD19
[LTl9], CD20 [2H7], CD56 [MEM-188] and CD83
[HBI5e] (Serotec, Oxford, United Kingdom) were used at
previously optimised titrations. The concentration of each
CB2 antibody was optimised by testing a range of titres from
1:100 to 1:1000, with a 1:400 dilution providing an optimal
signal for each CB2 antibody tested (data not shown).
Single cell suspensions were incubated simultaneously
with anti-CB2 and the respective fluorophore conjugated
primary antibodies on ice for 45 minutes. Unbound primary
antibody was removed by washing with PBS supplemented
with 1% foetal bovine serum. Anti-CB2 antibodies were
then detected with goat anti-rabbit Alexa 488 or 647
(Invitrogen, Carlsbad, CA) (detection of ABR, Sigma and
Abeam CB2) or sheep anti-goat Alexa 488 (Invitrogen)
(detection of Santa Cruz CB2). Secondary antibodies
were diluted 1:500 with PBS supplemented with 1% foetal
bovine serum and incubated for 20 minutes with cells on
ice. The activation status of monocytes was monitored
using CD83, and CD69 was used for B-lymphocytes and
T-lymphocytes (30-31). For all experiments a minimum of
3 donors were used and for each donor the experiment was
repeat at least three times.
 Int. J. ImmunopathoI. PharmacoI.
Two-colour live-cell immunocytochemistry
Live cell immunofluorescence was conducted by
staining the cells using the method described above
for flow-cytometry. Briefly, PBMC were probed with
antibodies detecting CD3 [UCHTl], CDI4 [MEM-18]
(Serotec) and CB2 (ABR), which were subsequently
detected with the corresponding isotype-specific goat anti-
mouse or goat anti-rabbit secondary antibodies conjugated
to Alexa 488 or 555 (Invitrogen). Cells were maintained at
4 T. Immediately prior to imaging, the cells were placed
on glass slides, cover slipped and allowed to settle for
approximately 2 minutes. Images were acquired using
a Leica DMR microscope equipped with epi fluorescent
filters for UV, FITC and TRITC. Images were acquired
using a 63x HCX PL Apo objective and a Nikon Digital
Sight DS-UI camera, and processed using EclipseNet
(Nikon) image acquisition software and Photoshop
(Adobe Systems, Mountain View, CA).
RESULTS
Comparison ofthe stainingpattern ofCB2 antibodies
in major leukocytes subsets
Four commercially available antibodies directed
at the N-terminus of CB2 were compared for
the detection of cell-surface CB2 receptors (Fig.
1). PBMC were stained to distinguish between
B-lymphocytes (CD20+), CD4+ T-lymphocytes
(CD4+/CD3+), CD8+ T-lymphocytes (CD8+/CD3+),
monocytes (CDI4+) and natural killer cells [NK,
(CD3neg/CD8mid/CDI6+/CD56+)] (Fig. 1, left hand
panels). The CB2 antibody comparison demonstrated
that the Santa Cruz CB2 antibody was not able
to detect cell surface CB2 in any of the leukocyte
subsets. The Sigma, ABR and Abeam CB2 antibody
staining indicated that some T-lymphocytes (CD4+
and CD8+) had low levels of cell surface CB2. This is
a significant observation as other studies employing
histology and western blotting have reported that
steady-state T-lymphocytes (21, 32) do not express
CB2. We found that CB2 protein expression by
NK cells, B-lymphocytes and monocytes was
considerably higher than that for T-lymphocytes. The
Sigma, Abeam and ABR CB2-antibodies produced a
relatively similar staining pattern for each cell type.
However, the Sigma antibody appeared to strongly
bind to a subset ofB-lymphocytes and NK cells; this
intense level of staining was not observed with the
other CB2 antibodies tested.
Whilst validating the staining profile of a
27
subsequent batch of the Abeam CB2 antibody
(same Cat#, different lots#) we noted a substantial
difference in the intensity of the staining (Fig. 2).
The staining profile of batch I was compared with
batch 2 using previously optimised titre conditions
determined for batch I (1 :400 dilution). Fig. 2
shows an example of the direct comparison of the
two Abeam batches used at 1:400 dilution on the
same PBMC preparation in the same experiment.
This revealed that the profile was similar however,
that the staining from Abeam batch I was an order of
magnitude stronger than that of the newer lot (batch
2) of Abeam CB2 antibody. This is depicted in the
figure by the difference in the rightward shift for
CB2 staining (black-line in the flow-chart) for each
leukocyte subset.
Detailed characterisation of CB2 expression by
human leukocyte subsets
Flow cytometry was conducted using PBMC
from 5 healthy volunteers to determine the relative
level of CB2 expression by CD8+ and CD4+ T-
lymphocytes, B-lymphocytes, monocytes and NK-
cells. All ofthese experiments were conducted using
the CB2 antibody obtained from ABR (Fig. 3).
A significant observation from this study is that
some steady-state T-lymphocytes express a low level
of CB2 receptor protein. The level of CB2 expressed
by T-lymphocytes was higher in some subjects than
others. This is demonstrated most clearly between
subject 2 [modest 5-fold change in mean fluorescent
intensity (MFI)] and subjects 3 and 4 (CB2 levels
were negligible), which may reflect the natural
variation in the expression of CB2 by different
individuals (Fig. 3a). Most B-lymphocytes, NK cells
and monocytes expressed moderate to high levels of
cell surface CB2. Interestingly, the highest levels
of cell surface CB2 were detected on NK cells for
subjects 2 and 4, whereas monocytes had the highest
level of CB2 for subjects 1 and 2. The variation in
CB2 expression for each cell type between subjects
is shown in Fig. 3b. CB2 expression by NK cells
showed the greatest degree of variation across
subjects ranging from a 5-fold to 25-fold shift for
subjects 3 and 4, respectively (Fig. 3b). Again,
this may represent an example of normal variation
which warrants further investigation. Such variation
makes ranking expression levels inappropriate as
28 E.S. GRAHAM ET AL.

CB2 CB2 CB2 CB2

Abcam ABR Santa Cruz Sigma

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lir;:::] 1 CD' '·,mp"oc,'"

I~C~~:j] I~LAJ ]
CD16 CD56
NK cell s

1 12\ ~ 1]
CD69
B·lymphocyl es

I'r -~~ I ---- - - - - - - - - - - - - ---1.-

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CD14
... ..r", ~
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CD83
Monocyl es

CB2 rece ptor

CD69 : indicates ecuvatco status of lymphocytes
CDS3 : Indica tes acnvatce statu s of monocytes
C016 and COS6 : provid e con formatIOn of NK cell pheno type

Fig. 1. Comparison of CB2 antibody staining of human PBMC using flow-cytometry. The expression of CB2 protein
was investigated using four N-terminal CB2 antibodies from Abeam, ABR, Santa Cruz and Sigma. Flow-cytometry
was conducted using the following cell-surface marker expression profiles to distinguish between leukocyte subsets;
CD4+/CD3+ for CD4+ T-Iymphocytes, CD8+/CD3+ for CD8+ T-Iymphocytes, CD3neg/CD8+/CDJ6+/CD56+ for NK
cells, CD20+ for B-Iymphocytes and CDJ4+ for monocytes. CD69 indicates activation status for lymphocytes; CD83
indicates activation status for monocytes. The left-most flow charts show the gated population for each leukocyte subset.
Representative cytometric expression profiles for CB2 are shownfor each cell type with each ofthe antibodies tested. The
control stain for each leukocyte population is indicated by the shaded grey area and the CB2-staining is indicated by the
black line. 3 donors were used and for each donor the experiment was repeated at least three times. Data shown is from
a single experiment.

il[ AJ]:~[Z[J >lOU ~[AJ

CD4 T-Iymphocytes CD8 T-Iymphocytes NK cell s B-Iymphocytes

Abcam ~ ~ ~ ~
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Batch 1
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CB2 receptor

Fig. 2. Comparison of the staining profiles of human CD4 and CD8 T-Iymphocytes, B-Iymphocytes, NK-cells and
monocytes using two different batches ofthe Abeam CB2 antibody. Flow-cytometry was conducted using human PBMC
and two distinct lots ofAbcam (ab356J) CB2 antibody to demonstrate the batch variability and importance ofappropriate
validation. The control stain for each leukocyte population is indicated by the shaded grey area and the CB2-staining is
indicated by the black line. Data shown isfrom a single experiment, which is representative ofthree donors.
 Int. J. Immunopathol. Pharmacol. 29

CD4 CDS
NK cells B·lymphocytes Monocytes

Su bject 3 J

Subject 4 J

CB2 receptor
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Fig. 3. Characterisation of CB2-expression by the major leukocyte subsets in PBMC from multiple healthy subjects.
a) The black line on the histograms show the relative expression levels of CB2 (ABR antibody) by CD4 and CD8 T-
lymphocytes, Bslymphocytes, monocytes and NK cells. The control stain for each leukocyte population is indicated by the
shaded grey area. CB2 expression was measured in five healthy subjects. b) The scatter plot shows the fold change in
the geometric mean fluorescent intensity (MFI) across subjects for each cell type; CD8' T-lymphocytes (CD8), CD4+ T-
lymphocytes (CD4), B-lymphocytes (B), NK-cells (NK) and monocytes (M). (c-j) Dualfluorescence immunocytochemistry
ofcell surface CB2 expression by monocytes and T-lymphocytes. Cell surface labelling ofthe CB2 receptor with the ABR
CB2 antibody shows expression by (c-d) monocytes (CD14+) and (e-j) T-lymphocytes (CD3'). For clarity the arrows
highlight the position ofthe T-lymphocyte in panel c and d. Scale bars = 10 um. The immunocytochemistry was repeated
with PBMC from 5 subjects.

our data suggest that for some subjects NK cells
have the highest level whereas for other individuals
monocytes express the highest. The low level of
CD69 (B and T lymphocytes) and CD83 (monocytes)
expression (see Fig. I; data not shown in subsequent
figures) demonstrates that the CB2 expression profile
measured in this study relates specifically to immune
cells under steady state conditions.
Live cell imaging of cell-surface eB2 receptor
expression
Flow cytometry results were verified by
immunocytochemistry using live cell imaging on
PBMC preparations. Dual immunofluorescence was
conducted on PBMC cultures with the ABR CB2
antibody (I :400) and markers for T-Iymphocytes
(CD3) and monocytes (CD14), which represent
30 E.S. G RA HAM ET A L.

(a) ~

FSC /SSC profile and

CD16 expression provide
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conformation of ,00 1(;' ,03 '0·
neutrophil phenotype ~
(b)
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(Collectivelythese markers
] SUbj ect 3 identify T and a·lymphocytes,
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10° 10' ,02 ,0" 1O·
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CB2 receptor

Fig. 4. Expression ofCB2 by neutrophils and whole-bloodfrom multiple healthy subjects. Flow cytometry was conducted
using whole blood fro m healthy subjects. a) Neutrophils were identified using their fo rward and side scatter profile and
cell surf ace expression of CD16 (33). CB2 expression was assessed using the ABR CB2 antibody, shown by the black
line with the grey area indicating the control stain. Data are f rom three subj ects repeated three time independently, data
shown is fro m one representative experiment. b) The major leukocyt e subsets account f or the maj ority of CB2-positive
cells present in human blood. Flow-cyt ometry was conducted using whole-blood from healthy donors stained with a
cocktail of F1TC-conjugated antibodies (CD3/CD 14/CD16/CD19/CD56) to simultaneously label lymphocytes, NK
cells, monocytes, and neutrophils. The absence of CB2+cells fro m the upper-left quadrant reveals that these leukocyte
populations account fo r the maj ority of the CB2-receptor expressing cells in human blood. These data are representative
ofthree independent experiments.

examples of CB2 Io\\ and CB2 high expressmg
cells, respectively. The immunocytochemistry
demonstrated that the monocyte CB2 levels were
sufficiently abundant to visualise by standard
immunocytochemistry, however the low level of
CB2 expressed by T-lymphocytes was not (Fig. 3
c-f).
Expression o/CE2 receptor by human neutrophils
The expression ofCB2 by human neutrophils was
investigated using whole-blood from health y donors
(PBMC preparations do not contain neutrophils due
to differential gradient separation). Importantly,
the blood was proce ssed immediately to minimise
the risk of acti vating the cells. Flow cytometry
was conducted using the ABR CB2 antibody and
neutrophils were ident ified on the basis of their
forward/side scatter profile and surface expression of
CD16 (33) (Fig. 4a) . Steady state human neutrophils
have moderate cell surface CB2 expression (Fig.
4a). As a final step in the characterisation, whole
blood was stained with markers inclusive for
T-lymphocytes (CD3 +), monocytes (CDI4+), B-
lymphocytes (CD 19+), neutrophils (CD 16+), and NK
cells (CD56+) to determine whether there were any
leukoc yte populations expressing CB2 not accounted
for (Fig. 4b ). The absence of a CB2 + population in
the upper left-quadrant of the flow-chart in Fig. 4b
indicates that there were no additional populations
expressing CB2 that were not already accounted
for.
DISCUSSION
We conducted a deta iled study to precisely define
the relati ve expre ssion of CB2 by blood-borne
 Int. J. Immnnopathol. Pharmacol.
immune cells from healthy human volunteers. This
is important as it will reveal those cells that are
potentially regulated by the endocannabinoid system
and those that represent direct targets for therapeutic
CB2-ligands.
The lack of specificity of GPCR receptor
antibodies (34) including CBl receptors (35) is an
ongoing challenge for researchers. We therefore
tested four different commercially available CB2
antibody preparations (from ABR, Abeam, Sigma
and Santa Cruz) on blood from healthy volunteers
to determine CB2 in living immune cells, The
antibodies (all recognise N-terminal epitopes of
CB2) were chosen primarily for their potential to
detect cell-surface CB2 in living immune cells. In
addition, the ABR CB2 (21, 23) and Abeam CB2
(36) antibodies have been used successfully to detect
endogenous CB2 receptors on human tissue sections
using immunohistochemistry.
Initial comparative experiments were conducted
using all four CB2 antibodies to assess the expression
patterns in PBMCs. The ABR, Abeam and Sigma
antibodies produced very similar CB2 detection
patterns in PBMC preparations, with moderate
to high levels of CB2 detected in monocytes, B-
lymphocytes, and NK cells in contrast to CD4+
and CD8+ T-lymphocytes that had very low levels
of CB2. These initial comparative studies were
particularly reassuring in that the staining pattern of
these three CB2 antibodies provided good agreement.
Furthermore, the striking difference in binding
between different leukocyte subsets, for example
the T-lymphocytes (CB2 1ow) and NK cells (CB2 high) ,
provided confidence that the CB2 antibodies were
binding genuine CB2 receptors. In contrast, the
Santa Cruz CB2 antibody failed to detect cell-surface
CB2 in any of the PBMC leukocytes, including B-
lymphocytes or NK cells which have been reported
to express the highest levels of CB2 mRNA (7).
While precise details are not available, the epitope
used to raise this CB2 antibody is located within
amino acids 10-60. If the IgG binds amino acids 33-
60 which comprise the first transmembrane domain
and intracellular loop, then this antibody is unlikely
to be suitable for live-labelling procedures. We have
used this antibody successfully for histological
localisation of CB2 in ischemic rodent brain
(25), implying suitability under certain detection
31
conditions. This finding stresses the importance
of full disclosure by commercial providers of the
epitope sequence to which the antibody is raised.
On closer scrutiny ofthe staining profile obtained
with the Sigma CB2 antibody (Fig. 1), we noted that
some NK cells and B-lymphocytes stained intensely
(beyond the 4 th log) which was not consistent with
a normal profile of expression. This high intensity
signal is more indicative of non-specific binding
and was not observed in the other cell types or for
the ABR or Abeam CB2 antibodies. Because of this
unusual staining pattern it was decided to exclude the
Sigma CB2 antibody from further experiments in this
study. We also noted that although a second batch of
the CB2 antibody purchased from Abeam produced
a similar profile of staining to our original batch the
staining intensity was approximately an order of
magnitude less. This demonstrates the importance
of batch consistency, in particular for human studies
or pathological tissues (clinical) where quantitative
assessments are being made. Consistent results
were obtained using the ABR antibody hence this
antibody was used for further analysis.
One of the most striking observations in this
study is that CB2 receptor protein was expressed by
all of the major leukocyte subsets present in human
blood (distinguishable with current cell surface
markers). In all 5 of our subjects the lowest level
of CB2 expression was by the T-lymphocytes. We
have demonstrated for the first time that steady-
state T-lymphocytes in some healthy individuals
express low levels of CB2 receptor protein. Our
data is consistent with there being detectible CB2
mRNA in both CD4+ and CD8+ T-lymphocytes (7)
and advances the findings of others (32), where CB2
expression was detected in activated T-lymphocytes,
but not steady state cells. This difference may
simply be an issue of greater sensitivity provided
by flow-cytometry over other methods (32). Further
investigation is required to determine whether the
low level of CB2 expression we observed in steady
state T-lymphocytes is functionally relevant and
whether it is sufficient to confer cannabinoid control.
Of the other major leukocyte subsets present in
healthy human blood (i.e. B-Iymphocytes, NK cells,
monocytes and neutrophils) all had abundant levels
of CB2 receptor protein. These data indicate that
all of the major human leukocyte subsets express
32 E.S. GRAHAM ET AL.
CB2 protein and are therefore likely regulated by
the endocannabinoid system. Furthermore, it is
important to recognise that each of these cell types is
therefore a potential target for CB2-mimetic drugs.
Earlier studies assessing CB2 expression in
humans have primarily focused on lymph nodes;
lymph nodes are a crucial part of the immune
system where immune responses are initiated. Using
immunohistochemistry Galiegue and colleagues
detected CB2 receptor in human tonsils (7), noting
CB2-immunoreactivity in B-lymphocyte rich areas.
Similarly, another study detected CB2 expression
by B-lymphocytes in both healthy and tumour-
infiltrated lymph node sections (21), however,
interestingly, T-lymphocytes lacked CB2 expression.
In contrast, in T-non-Hodgkin's lymphoma lymph
nodes T-lymphocytes did express CB2 (21).
Although some paralells can be drawn between cells
which express CB2 in the lymph node and blood
i.e. B-lymphocytes, the expression levels of CB2
by immune cells in lymph nodes under steady state
or reactive conditions may not be representative of
immune cells in blood, highlighting the importance
of assessing CB2 expression in each individual
tissue. This study represents the first comprehensive
analysis detailing CB2 protein expression by each of
the major human leukocyte subsets in whole blood
and PBMC from healthy individuals.
ACKNOWLEDGEMENTS
The authors wish to thank all donors for
participating in this study. This research was funded
by the New Zealand Marsden Fund, Auckland
Medical Research Foundation and an Auckland
University Early Career grant awarded to ESG.
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